Poly(A) Polymerase Tailing Kit对单链RNA添加Poly(A)尾的效果图。在50μl反应体系中,加入5 μg 541nt的单链RNA,以及图中指定量的Poly(A) Polymerase,37ºC孵育30 min,65ºC孵育20min终止反应。以下为琼脂糖凝胶电泳检测效果图。
Lane 1阴性对照(未加Poly(A) Polymerase);
Lane 2 0.008 U PAP;
Lane 3 0.04 U PAP;
Lane 4 0.2 U PAP;
Lane 5 1 U PAP;
Lane 6 5 U PAP;
Lane 7 10 U PAP;
Product Details
Product Details
Product Specification
| Stability & Storage | Store at -25 ~ -15℃ for 2 years |
| Reference |
Components
组分 |
UA070116-50 Rxns |
UA070116-250 Rxns |
|
E. coli Poly(A) Polymerase (5 U/µl) |
100 μl |
5 × 100 μl |
10 X PAP Reaction Buffer |
300 μl |
5 × 300 μl |
10 mM ATP |
300 μl |
5 × 300 μl |
RNase Free dH2O |
1 ml |
5 × 1 ml |
Protocol
1. Prepare the following reaction system on ice according to the table below:
Component |
Volume |
Final Concentration |
10 X PAP Reaction Buffer |
5 μl |
1 X |
RNase Inhibitor (40U/μl) |
1.25 μl |
1 U/μl (Optional) |
|
E.coli Poly(A) Polymerase (5 U/μl) |
2 μl |
0.2 U/μl |
10 mM ATP |
5 μl |
1 mM |
RNase Free dH2O |
Up to 50 μl |
- |
Guidelines
1. Due to the involvement of RNA manipulation, strict adherence to RNA operation protocols is required to avoid RNase contamination. Related reagents and consumables must be treated with DEPC to remove RNase or ensured to be RNase-free. 2. The RNA used for the tailing reaction should be appropriately purified before use and dissolved in RNase-free dH2O. The solution should not contain EDTA or salts. 3. If it is difficult to ensure a strictly RNase-free environment, it is recommended to add an appropriate amount of RNase Inhibitor to the reaction system to enhance the stability of RNA in the solution. 4. During experimental operations, enzyme products should be kept on ice at all times and stored at -20°C immediately after the experiment.
Picture
Picture
Bioactivity
